Testcrosses are an efficient strategy for identifying cis regulatory variation: Bayesian analysis of allele specific expression (BASE)

Miller, B.; Morse, A.; Borgert, J. E.; Liu, Z.; Sinclair, K.; Gamble, G.; Zou, F.; Newman, J.; Leon-Novelo, L.; Marroni, F.; McIntyre, L.

doi:10.1101/2020.10.01.322362

bioRxiv · 10.1101/2020.10.01.322362

Testcrosses are an efficient strategy for identifying cis regulatory variation: Bayesian analysis of allele specific expression (BASE)

Miller, B.·Morse, A.·Borgert, J. E.·Liu, Z.·Sinclair, K.·Gamble, G.·Zou, F.·Newman, J.·Leon-Novelo, L.·Marroni, F.·McIntyre, L.

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Abstract

Allelic imbalance (AI) occurs when alleles in a diploid individual are differentially expressed and indicates cis acting regulatory variation. What is the distribution of allelic effects in a natural population? Are all alleles the same? Are all alleles distinct? Tests of allelic effect are performed by crossing individuals and comparing expression between alleles directly in the F1. However, a crossing scheme that compares alleles pairwise is a prohibitive cost for more than a handful of alleles as the number of crosses is at least (n2-n)/2 where n is the number of alleles. We show here that a testcross design followed by a hypothesis test of AI between testcrosses can be used to infer differences between non-tester alleles, allowing n alleles to be compared with n crosses. Using a mouse dataset where both testcrosses and direct comparisons have been performed, we show that [~]75% of the predicted differences between non-tester alleles are validated in a background of [~]10% differences in AI. The testing for AI involves several complex bioinformatics steps. BASE is a complete bioinformatics pipeline that incorporates state-of-the-art error reduction techniques and a flexible Bayesian approach to estimating AI and formally comparing levels of AI between conditions. The modular structure of BASE has been packaged in Galaxy, made available in Nextflow and sbatch. (https://github.com/McIntyre-Lab/BASE_2020). In the mouse data, the direct test identifies more cis effects than the testcross. Cis-by-trans interactions with trans-acting factors on the X contributing to observed cis effects in autosomal genes in the direct cross remains a possible explanation for the discrepancy.

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Miller, B., Morse, A., Borgert, J. E., Liu, Z., Sinclair, K., Gamble, G., Zou, F., Newman, J., Leon-Novelo, L., Marroni, F., McIntyre, L.. 2020-10-03. Testcrosses are an efficient strategy for identifying cis regulatory variation: Bayesian analysis of allele specific expression (BASE). https://doi.org/10.1101/2020.10.01.322362

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